体外扩增CIK细胞对结肠癌SW480细胞株杀伤作用的研究  

KILLING EFFECT OF CYTOKINE-INDUCED KILL CELLS ON COLON CANCER LINES SW-480 IN VITRO

作　　者：池小钜[1] 刘汉锋[1] 蓝东[1] 

机构地区：[1]广西医科大学第一附属医院,南宁530021

出　　处：《广西医科大学学报》2014年第3期372-377,共6页Journal of Guangxi Medical University

基　　金：广西自然科学基金资助项目(No.桂科自03201208022)

摘　　要：目的：观察细胞因子诱导的杀伤细胞（CIK细胞）体外增值情况,了解其扩增后杀伤结肠癌SW480细胞株最佳比例及最有效的维持时间.方法：分离健康人外周血单个核细胞在体外诱导制备成CIK细胞,计数不同培养时间的CIK细胞,采用流式细胞仪检测其表型特征.倒置显微镜下观察CIK细胞作用与SW480结肠癌细胞的形态学变化,采用HE染色、AO/EO荧光染色和MTT法检测CIK细胞对SW480结肠癌细胞株的杀伤活性.结果：CIK细胞在体外培养14 d细胞增殖倍数为（100.22±8.68）倍,培养21 d细胞增殖倍数为（300.86±10.13）倍,两者比较差异有统计学意义（P＜0.05）;培养28 d细胞增殖倍数为（305.06±5.13）倍,与培养21 d的细胞增殖倍数比较差异无统计学意义（P＞0.05）.培养21 d的CIK细胞中CD3＋、CD56＋双阳性细胞的百分比为（45.3±5.1）％,与28 d相比较差异无统计学意义（P＞0.05）,第35天细胞增殖（212.50±4.25）倍,呈下降趋势.CIK细胞对结肠癌SW480细胞株有明显的杀伤作用,最高杀伤率为（68.21±1.31）％.结论：CIK细胞具有较强的抗结肠癌细胞活性,体外培养14～21 d具有较强的抗瘤活性,此时应用较为合适.CIK细胞作用于肿瘤细胞应达到一定数量并维持一定时间,可达到较好的抗瘤活性.Objective：To observe the killing effect of cytokine-induced kill cells （CIK） from healthy donor on colon cancer cell lines SW480 in vitro. Methods： Human peripheral blood mononuclear cells from healthy donors were induced to become CIK cells by cytokine in vitro incubation. The proliferation ability of CIK cells was measured by calculation in different culturing time and the phenotype of CIK cells was analyzed by a flow cytometer. Inverted microscope to observe morphological changes of SW480 colon cancer after added the CIK cells. The antitumor cytotoxicity of CIK cells against SW480 cell lines was measured by HE staining, AO/EO double fluorescent staining and MTT assay. Results： After 14 days in the culture, the cells expanded to median of （100.22±8. 68） times more they used to be, while comparing to the statistics acquired on the 21th day, which showed a median of （300.86±10. 13） times, the data was different statistics （ P 〈0.05）. However, the results from the 28th day which showed a median of （305.06±5.13） times, showed no significant difference, comparing with the data on the 21th day. The number of CD3＋ CD56＋ cells had a significant increase in the percentage rate. The percentage was to （45.3±5.1）% after 21 days culturing. The CIK cells possessed significant ability to kill SW480 colon cancer cells in vitro, the maximum killing efficiency is about （68.21±1.31）%. Conclusion： CIK cells own strong anti cancer activity against sw480 colon cancer cells in vitro. The suitable time for clinical application is on days 14 to 21, which is the proliferation peak. The effect of CIK cells on tumor cells should reach a certain number and maintain for a period of time, the possibility of better antitumor activity reached.

关 键 词：CIK细胞 结肠癌SW480 HE染色 AO EO荧光染色 MTT法检测 

分 类 号：R735.35[医药卫生—肿瘤]